α-Glucosidase​

UNIT DEFINITION

One unit (U) is defined as the amount of enzyme which produces 1 μmol of PNP per min under the conditions described below.
CAS # 9001-42-7

REAGENTS PREPARATION

Reagent I: 100 mM PBS, pH 7.0.

Reagent II: 20 mM p-Nitrophenyl-α-D-glucopyranoside (PNPG).

Reagent III: 200 mM Na2CO3.

Reagent IV: 200 mM, pH 7.0 potassium

phosphate buffer, contain 0.05% Tween 20 and 1 mM EDTA.

Sample: Dilute the sample to 0.006-0.022 U/mL by Reagent IV.

PROCEDURE

1.   Add 1mL Reagent I, 0.5 mL Reagent II to a test tube, and preincubate for 5 min at 37 ℃.

2.   Add 0.5 mL sample and mix, react for 15 min in 37 ℃, then add 2 mL reagent III to stop the reaction.

3.  Record the ΔAs at 400 nm in a spectrophotometer.

4.  Exchange the adding order of reagent III and sample, and measure the blank absorbance ΔAb.

∆A = ∆As – ∆Ab